Hypocholesterolemic compounds

ABSTRACT

BENZYL-SUBSTITUTED CYCLOOCTANONE COMPOUNDS WHICH EXHIBIT HYPOCHOLESTEROLEMIC PROPERTIES.

"United States Patent 3,781,444 HYPOCHOLESTEROLEMIC COMPOUNDS Bobby R. Ezzell, Houston, Tex., Eugene R. Fluck,

Berwyn, Pa., and Louis R. Haefele, Winston-Salem, N.C., assignors to R. J. Reynolds Tobacco Company, Winston-Salem, NC. NoDrawing. Continuation-impart of abandoned application Ser. No. 105,076, Jan. 8, 1971. This application 1 Nov. 12, 1971, Ser. No. 198,445 Int. Cl. A61k 27/00 US. Cl. 424-331 3 Claims ABSTRACT OF THE DISCLOSURE Benzyl-substituted cyclooctanone compounds which eX- hibit hypocholesterolemic properties.

,v This application is a continuation-impart of our copending application Ser. No. 105,076 which was filed J an. 8,

1971, now abandoned.

N This invention relates to new compositions of matter having useful pharmacological properties.

Elevated levels of cholesterol in the blood stream are believed to play a significant role with respect to cardiovascular disease' and particularly atherosclerosis. In the treatment and prevention of atherosclerosis it is desired to maintain normal blood cholesterol levels and/or to reduce blood cholesterol levels in hypercholesterolemic patients. It has now been found that chemical compounds having structural formulae I and II exhibit hypocholesterolemic properties:

CH2) (Formula 1 where X=O, NOH, NNH2, NNHCH3, NNHC H NN(CH3)2 or R3 being H, OH, OCH3, OC2H5, CH3 OI' C2H5 and the aromatic ring being unsubstituted or substituted with not more than three substituents selected from C1, F, CN, NH CH OH, lower alkyl, lower alkoxy, lower alkylamino and lower dialkylarnino, and

with R being H, OH, OCH OC H CH or C H and the aromatic ring in each of R and R being independently unsubstituted or substituted with not more than three substituents selected from C1, F, CN, NH 0P OH, lower alkyl, lower alkoxy, lower alkylamino and lower dialkylamino.

3,781,444 Patented Dec. 25, 1973 The term lower as used herein in connection with alkyl or alkoxy groups refers to groups having from 1 to 5 carbon atoms.

Compositions which are particularly preferred in accordance with the present invention include compounds embraced by structural Formulae III and IV:

& 0 z) a where X=O, NOH, NNHZ, NNHCH3, NNHC6H5, mucu or NNHCONH2, R1:

(Formula III) with R being H, OH, OCH OC H CH 0r C H and the aromatic ring being unsubstituted or substituted with not more than three substituents selected from C1, F, CN, NH CB OH, lower alkyl, lower alkoxy, lower alkylamino and lower dialkylamiuo provided that when X=O and the aromatic ring in R is unsubstituted, R is other than H or OH, and

t C R Q1 N\ (Formula IV) where R and R R3 being H, OH, OCH3, 0C2H5, 01' C2H5 and the aromatic ring in each of R and R being independently unsubstituted or substituted with not more than three substituents selected from C1, F, CN, NH CF OH, lower alkyl, lower alkoxy, lower alkylamino and lower dialkylamino.

Preparation of the novel compounds of the invention is illustrated by the following examples.

EXAMPLE I 2- (p-chlorobenzyl cyclooctanone,

2- (2,6-dich1orobenzyl) cyclooctanone,

2- (m-fluorobenzyl) cyclooctanone,

2- (2-fluoro-6-chlorobenzyl cyclooctanone, 2- (o-methylbenzyl) cyclooctanone,

2- (m-trifluoromethylb enzyl cyclooctanone, 2- (p-methoxybenzyl cyclooctanone, 2-(2,4-dimethoxybenzyl cyclooctanone,

2- (2,4,5 -trimethoxyb enzyl) cyclooctanone,

2-(3,4-dihydroxybenzyl)cyclooctanone, 2-(Z-hydroxy-S-methoxybenzyl)cyclooctanone,

2- (m-cyanob enzyl) cyclooctanone and 2- (2-chloro-4-dimethylaminobenzyl) cyclooctanone.

The oxime of Z-benzylcyclooctanone is prepared by placing an 18.5-gram portion of Z-benzylcyclooctanone as obtained above in 150 milliliters of a 1:1 mixture of ethanol and pyridine containing 15 grams of hydroxylamine hydrochloride. The resulting mixture is heated for 3 hours at reflux temperatures. Most of the solvent mixture is then removed under reduced pressure and the residual oil is washed with water before dissolving in ether and washing the ether solution with several additional portions of water. Removal of the ether yields a solid which upon recrystallization from methanol gives crystalline 2-benzylcyclooctanone oxime, melting point 117-119" C. Also prepared in a similar manner are the oxime derivatives of 2- (o-chlorobenzyl) cyclooctanone,

2- (p-fluorohenzyl cyclooctanone,

2- (m-methylbenzyl) cyclooctanone,

2- (2,3 -dicllorobenzyl) cyclooctanone,

2- (o-trifluorornethylb enzyl) cyclooctanone,

2- (o-methoxybenzyl) cycloo ctanone,

2- (2, 3 -dimethoxyb enzyl) cyclooctanone,

2- p-hydroxybenzyl cyclooctanone,

2- 3-ethyl-4-hydroxy-S-isopropylbenzyl cycloo ctanone, 2- (2-hydroxy-5-methoxybenzyl) cyclooctanone and 2- (p-dimethylaminobe-nzyl) cyclooctanone.

The hydrazones, methylhydrazones, N,N-di1nethylhydrazones, phenylhydrazones and semicarbazones of the 2-benzylcyclooctanones are prepared using the general procedures described by L. F. Fieser and M. Fieser in Reagents for Organic Synthesis, John Wiley and Sons, Inc., New York, N.Y., 1967, especially pages 290, 436, 838 and 1000. Compounds converted to the hydrazone, methylhydrazone, N,N-dimethylhydrazone, phenylhydrazone and semicarbazone derivatives using these procedures are EXAMPLE II 2,8-dibenzylcyclooctanone is prepared by stirring 25 grams of cyclooctanone, 54 grams of benzaldehyde, 30 grams of potassium hydroxide and 500 milliliters of methanol at reflux temperatures for 4 hours. The mixture is then cooled to room temperature and 200 milliliters of water are added followed by 300 milliliters of ether. The ether layer is separated and the ether and methanol are then removed by distillation. The residue is taken up in 200 milliliters of ethanol and hydrogenated at 1,000 p.s.i.g.

and ambient temperature using a percent palladium on carbon catalyst. The catalyst is removed by filtration 7 0 and the solvent is removed by distillation. Recrystallization of the crude product is effected by dissolving in 150 milliliters of methanol and seeding with crystalline 2,8-dibenzylcyclooctanone (M.P. -77" C.). Cooling of the seeded methanolic solution gives 16 grams of crystals 4 'with melting point 60-67" of this material from methanol gives one isomer of 2,8- dibenzylcyclooctanone, melting point 75-77 C., as a first crop and a second isomer of 2,S-dibenzylcycloOctanone, melting point 66-68 C., as a second crop.

EXAMPLE III A mixture consisting of cyclooctanone (12.5 grams), benzaldehyde (27 grams), potassium hydroxide (30 grams) and 200 milliliters of methanol is stirred at reflux temperatures for 4 hours and is allowed to stand overnight. The crystalline precipitate which forms is collected by filtration and is washed with hot methanol to give 17 grams of 2 benzylidene-8-(u-rnethoxybenzyl)cyclooctanone, melting point 136-139 C. Hydrogenation of a 10 gram portion of this material at 60 p.s.i.g. in'300 milliliters of ethanol using 10 percent palladium on carbon as the catalyst gives 3.9 grams of 2-benzyl-8-(ot-methoXy-,

benzyl)cyclooctanone, melting point 104106 C., as a first crop of crystals from ethanol. Removal of a second crop of crystals and concentration of the mother liquor yields 1.1 grams of an isomeric 2-benzyl-8-(a-meth0xybenzyl)cyclooctanone, melting point 83-84 C.

The above described procedure is also used in preparing EXAMPLE IV An alternative procedure for preparing 2,8-dibenzylcyclooctanones involves the corresponding 2,8-dibenzylidenecyclooctanones as the intermediates. Thus, a mixture comprising 12.5 grams of cyclooctanone, 27 grams of benzaldehyde, 15 grams of potassium hydroxide and 300 milliliters of methanol is stirred at reflux temperatures for 4 hours. To the hot, stirred mixture are slowly added 400 milliters of -t-butyl alcohol with simultaneous removal of the methanol by distillation. The mixture is then cooled, water is added and the product is extracted with ether. The crude product obtained from the ether extract is chromatographed on activated alumina to give 2,8-dibenzylidenecyclooctanone. This intermediate is hydrogenated at '60 p.s.i.g. in ethanol using 5 percent palladium on carbon. Fractional crystallization of the hydrogenation product from methanol affords the isomeric 2,8-dibenzylcyclooctanones, M.P. 75-77 C. and 66-68 C.

The foregoing procedure is also used for preparing 2,8-'bis(p-chlorobenzyl) cyclooctanone,

2, S-bis 3 ,5 -dichlorobenzyl) cyclooctanone, 2,8-bis (o-fluorobenzyl) cyclooctanone, 2,8-bis(2 fluoro-6-chlorobenzyl cyclooctanone, I 2,8-bis o-methylbenzyl) cyclooctanone,

2,8-bis (m-trifiuoromethylbenzyl) cyclooctanone, 2,8-bis (p-butylbenzyl) cyclooctanone,

2,8-bis (p-methoxybenzyl) cyclooctanone C. A second recrystallization.

EXAMPLE V 2,8-disubstituted cyclooctanonesyvhich are unsymmetrically substituted are prepared from 2-benzylcyc1ooctanones which have been previouslyiprepared by the procedure of Example I or by the procedure for Z-benzylcyclooctanone as described by R. P. Houghton and E. S. Waight in J. Chem. Soc., 978-981 (1969). The particular Z-benzylcyclooctanone is placed in a methanolic solution of the desired aromatic aldehyde'and potassium hydroxide and the resulting mixture is stirred at reflux temperatures for a. period of time, The crude product is recoveed by ether extraction and subjected to hydrogenation at 60 p.s.i.g. in ethanol using percent palladium on carbon as the catalyst to give the unsymmetrical 2,8-disubstituted cyclooctanones.

Compounds prepared by the foregoing procedure are EXAMPLE v1 1 2-benzylidenecyclooctanones are prepared according to the procedure of Braude et al. as described in J. Chem. Soc.,- 4711-4719 (1957). .The 2-benzylidenecyclooctanones arethen subjected to condensation reaction conditions (such as methanolic potassium hydroxide) in the presence of equimolar amounts of Ta desired benzaldehyde to give substituted-benzyl 2-benzylidene-8-(a-alkoxybenzyl)cyc1ooctanones. Hydrogenation of these intermediates at 60 p.s.i.g. in thepresence of 5 percent palladium on carbon gives the corresponding 2-benzyl-8-(a-alkoxybenzyl)cyclooctanones wherein the substitution on the aromatic rings is difierent. Compounds prepared by this procedure are 2-b enzyl-8- p-chloro-a-rnethoxyb enzyl) cycloo ctanone,

2- (p-chlorobenzyl) -8- a-methoxybenzyl) cyclooctanone,

2- (3,4-dichlorobenzyl) -8- (m-chloro-a-methoxybenzyl) cyclooctanone,

2- (2-fluoro-6-chlorobenzyl) -8- (o-fiuoro-a-methoxyb enzyl) cyclooctanone,

2- (p-methylbenzyl) -8- a-methoxybenzyl cyclooctanone,

2- (m-trifiuoromethyl -8- (pmethyl-a-methoxybenzyl) cyclooctanone,

2- (o-methoxybenzyl) -8- (p,a-dimethoxybenzyl) cyclooctanone,

2- (3 ,4-dimethoxybenzyl) -8- (p-hydroxy-u-methoxybenzyl) cyclooctanone,

2- (2,4-diethoxy-5-ethylbenzyl) 8- (m-ethyl-a-methoxybenzyl) cyclooctanone,

2- (2-hydroxy-3 -methoxybenzyl) -8- (m-chloroa-methoxybenzyl) cyclooctanone,

2- (3 ,5 -dirn ethoxy-4-hydroxybenzyl) -8- (3 ,4,5,a-tetramethoxybenzyl)cyclooctanone,

6 2- (p-cyanobenzyl) -8- (m-fluoro-u-methoxyb enzyl) cyclooctanone, j g 2- (p-diethylaminobenzyl) 8- m-cyano-a-methoxybenzyl) cyclooctanone, 2-(2-chloro-4-dimethylamino)-8-(p-chloro-a-methoxybenzyl)cyclooctanone, and 2-(p-chlorobenzyl)-8-(2-chloro-4-dimethylamino-umethoxybenzyl cyclooctanone.

EXAMPLE VII This example illustrates the preparation of benzylcyclooctanones wherein the aromatic ring is substituted with an amino group. 2-benzylcyclooctanone is allowed to react With equimolar amounts of p-nitrobenzaldehyde in refluxing methanolic potassium hydroxide for 3 hours. The 2-benzyl-8-(p-nitrobenzylidene)cyclooctanone which forms is removed by filtration and subjected to hydrogenation at 50 p.s.i.g. in thepresence of 10 percent palladium on carbon using a solvent system comprising equal portions of ethyl acetate and acetic acid. The 2-benzyl- S-(p-aminobenzyl)cyclooctanone which is obtained from the hydrogenation is purified by converting it to the hydrochloride salt by means of hydrogen chloride introduced into an ethereal solution of the crude product. The melting point of the crude 2-benzyl-8-(p-aminobenzyl) cyclooctanone hydrochloride is -195 C.

In a similar manner are prepared The preparation of (m-hydroxybenzyl)cyclooctanones is etfected using the general procedure of Braude et al. in J. Chem. Soc., 4717 (1957) for making 2-(a-hydroxybenzyl)cyclooctanone. For the preparation of 2,8-disubstituted cyclooctanones such as 2-benzyl-8-(a-hydroxy benzyl)cyclooctanone, low pressure hydrogenation in the presence of palladium on carbon is effective in reducing the olefinic bond in 2-benzylidene-8-(a-hydroxybenzyl) cyclooctanone which has been previously reported by R. Cornubert et al. in Bull. soc. chim. 5, 1493 (1938). Using the basic procedures mentioned, the following compounds are prepared:

2- (3 -fiuoro-4-chloro-a-hydroxybenzyl) cyclooctanone,

2- (3-fluoro-4-ethoxy-a-hydroxybenzyl cyclooctanone,

2- (p-dimethylamino-a-hydroxybenzyl) cyclooctanone,

2- (2,4-dichlorobenzyl) -8- 2,4-dichloro-u-hydroxybenzyl) cyclooctanone, V

2- (p-trifluoromethylbenzyl) -'8- (p-trifiuoromethylochydroxybenzyl) cyclooctanone,

2- (p-isopropylbenzyl) -8- (p-isopropyl-u-hydroxybenzyl) cyclooctanone, and

2- (3 -pyridylrnethyl) -8- (a-hydroxy-3-pyridylmethy1) cyclooctanone.

The hypocholesterolemic properties of the new compounds of the invention are shown by the following test data obtained by orally administering aqueous suspensions of the compounds to rats for a period of five days. The animals were fasted overnight prior to the fifth day of the experiment and the blood samples were obtained via cardiac puncture two hours after the last dnlg administration. The blood samples were then analyzed for total cholesterol.

The ability of compounds of this invention to lower serum cholesterol levels in the rat is seen from the above data. In addition to the ability to lower serum cholesterol levels, hypocholesterolemic agents desirably should not produce an accumulation of cholesterol intermediates such as desmosterol which are in themselves atherogenic. In this connection, a particularly preferred compound of this invention was compared with {ethyl ot-(p chlorophenoxy)isobutyrate (CPIB)], a drug in current clinical use. This involved a comparison of the hepatic cholesterol synthesis in animals. The parameters of special interest were alterations in cholesterol synthesis as measured by in vitro incorporation of C -labelled mevalonic acid into various lipid classes (including cholesterol and cholesterol esters) in liver slices taken from drug-treated animals and by accumulation of desmosterol or other atherogenie sterol intermediates.

Three randomly selected groups of rats weighing approximately 150 grams each were fed ad libitum throughout the experiment. Animals were given daily doses of an aqueous suspension of the appropriate drug or water orally by hypodermic syringe.

Group A-Water only Group BCPIB, 200 mg./kg. daily Group C--2,8-dibenzylcyclooctanone (MP. 75-77 C.),

100 mg./ kg. daily At appropriate intervals over a period of 28 days, one rat from each group was sacrificed and liver slices were prepared in duplicate for incubation with mevalonic-Z- C acid. The degree of incorporation of mevalonic-2-C acid into the various lipid classes present in the liver tissue was determined and the data indicated that there was no gross inhibition in the biosynthesis of either cholesterol or cholesterol esters in the Group B and Group C animals as compared with the control group. Moreover, there was no accumulation of sterol intermediates such as desmosterol apparent in the Group C animals relative to the control group. i

The compounds of this invention can be used as hypocholesterolemic agents at relatively low dosage levels. In

general, they can be employed in amounts" ranging from about 1.0 to 500 trig/kg. of the hostbody weighL'As' can be seen from the data in Table I, hypocholesterole'mic activity is exhibited by each of the 2,8-dibenzylcyclooctanone isomers isolated as well as by each of the 2-benzyl-8-(a methoxybenzyl)cyclooctanoneisomers obtained. It is preferred that the compoundsbe administered orally, but it is apparent that other meansfof administration may: be

equally effective. When given orally, the compounds can be administered in tabl et'form or as aqueous suspensions or emulsions. Compounds which are capable of forming acid addition salts may. be, administered in that form pro vided, of course, that therapeutically acceptable salts are used. Thus, for example, aminobenzyl and pyridylmethylsubstituted cyclooctanones may be converted to the, corresponding hydrochloride, sulfate, phosphate, acetate;

tartrate or gluconate salts and used in that form.

Those modifications and equivalents which fall within the spirit of the invention are to be considered a part thereof.

What is claimed is:

1. A process of lowering elevated serum cholesterol levels in a mammal in need thereof which comprises administering to said mammal in an amount sufficient to achieve a hypocholesterolemic effect a compound having the formula: t

where R and R I References Cited Chemical Abstracts, 63: 13126 (1965).

Chemical Abstracts 67:607032 (1967). Chemical Abstracts 69:96072y (1968).

ALBERT T. MEYERS,'P ri1nary Examiner N. A. DREZIN, Assistant Examiner U.S."'Cl. X.R. 424-263, 304, 321' UNITED STATES PATENT CFFICE CERTIFICATE OF CORRECTION Dated December 25, 1973 Patent No. 3 781,444

Inventor(s) p It is certified that error appears in the above-identified patent and that said Letters Patent are hereby corrected as shown below:

Column 2, lines 30-35, Formula IV should read as follows:

Column 3, line 60, "2,S-dibenzylcyclooctanone"should be 2,8-Dibenzylcyclooctano e Column 5, line'8", "2 8'-disubstituted" s'hou'ldbe' 2,8-

Disubstituted r A line 44, "2,8-benzylidenecyclooctanones" should be 2,8-Benzylidenecyclooctanones Column 6, line 13,- "Z-benzylcyclooctanone should-be v Z-Benzylcyclooctanone Column 7, third line from the bottom in Table I, after "84" insert C Signed and sealed this 9thday of April 197A.

(SEAL) Attest:

"c. MARSHALL DANN EDWARD I LFLEICHERJR.

Commissioner of Patents Attesting Officer USCOMM-DC 60376-P69 FORM PO-IOSO (10-69) 

